Scholarlink

Abstract <p><i>Aurantiochytrium</i> <i>limacinum</i> holds great promise for producing sustainable single-cell oil as an alternative to fish oil. However, research into its complex biological and biochemical characteristics and efforts toward strain improvement have been hampered by insufficient genetic tools. Until now, genetic transformations of <i>A. limacinum</i> have relied solely on chromosome integration, which is inefficient and prone to insertional mutagenesis and other issues related to genetically modified organisms (GMOs). This paper describes the first centromeric plasmid for <i>A. limacinum</i>. Amplification of this shuttle vector by <i>E. coli</i> enables direct delivery into <i>A. limacinum</i> via electroporation, where it undergoes stable replication and segregation into daughter cells. The key to the stable plasmid maintenance lies in a 500&#xa0;bp segment derived from chromosome 24 of <i>Phaeodactylum tricornutum</i>. While this segment does not significantly enhance the efficiency of vector transformation, it enables the replication and maintenance of the shuttle vector in the host cell as closed circular DNA. The plasmid from three transformants demonstrates a high segregation efficiency of 96.8 ± 0.3% (<i>n</i> = 3), even in the absence of antibiotic selection. This novel centromeric plasmid considerably enhances the flexibility of genetic manipulations and gene expression in <i>A. limacinum</i>, opening new avenues for its study and industrial application.</p> Key points <p><i>• First centromeric plasmid developed for genetic transformation in A. limacinum.</i></p> <p><i>• The novel plasmid enhances flexibility in genetic manipulation and gene expression.</i></p> <p><i>• The plasmid achieves 96.8 ± 0.3% (n = 3) segregation efficiency without antibiotic selection.</i></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Novel centromeric plasmid for stable extrachromosomal gene expression in Aurantiochytrium limacinum

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年7月4日

查看全文
  • Person Pesona Renta,
  • Cian-Huei Syu,
  • Ta-Yu Huang,
  • Yi-Ting Chang,
  • Yu-Feng Liang,
  • Ssu-Ting Chen,
  • Po-Wei Weng,
  • Ming-Chen Hsu,
  • Keng-Hung Lin,
  • Tsunglin Liu,
  • Anna C.-C. Jang,
  • Che-Chia Tsao,
  • Han-Jia Lin,
  • Hung-Yun Lin,
  • Yi-Min Chen

摘要

Abstract

Aurantiochytrium limacinum holds great promise for producing sustainable single-cell oil as an alternative to fish oil. However, research into its complex biological and biochemical characteristics and efforts toward strain improvement have been hampered by insufficient genetic tools. Until now, genetic transformations of A. limacinum have relied solely on chromosome integration, which is inefficient and prone to insertional mutagenesis and other issues related to genetically modified organisms (GMOs). This paper describes the first centromeric plasmid for A. limacinum. Amplification of this shuttle vector by E. coli enables direct delivery into A. limacinum via electroporation, where it undergoes stable replication and segregation into daughter cells. The key to the stable plasmid maintenance lies in a 500 bp segment derived from chromosome 24 of Phaeodactylum tricornutum. While this segment does not significantly enhance the efficiency of vector transformation, it enables the replication and maintenance of the shuttle vector in the host cell as closed circular DNA. The plasmid from three transformants demonstrates a high segregation efficiency of 96.8 ± 0.3% (n = 3), even in the absence of antibiotic selection. This novel centromeric plasmid considerably enhances the flexibility of genetic manipulations and gene expression in A. limacinum, opening new avenues for its study and industrial application.

Key points

• First centromeric plasmid developed for genetic transformation in A. limacinum.

• The novel plasmid enhances flexibility in genetic manipulation and gene expression.

• The plasmid achieves 96.8 ± 0.3% (n = 3) segregation efficiency without antibiotic selection.